Subcellular mapping of living cells via synchrotron microFTIR and ZnS hemispheres.
Subcellular mapping of living cells via synchrotron microFTIR and ZnS hemispheres.
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DOI：
10.1007/s00216-018-1245-x
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发表时间：
2018-10
影响因子：
4.3
通讯作者：
Cinque G
中科院分区：
文献类型：
--
作者：
Chan KLA;Fale PLV;Atharawi A;Wehbe K;Cinque G
FTIR成像是一种无标记、非破坏性的方法，在研究活细胞的生物过程中具有重要的应用价值。然而，水的长波长/低空间分辨率和强吸光度仍然是制约红外显微技术在体外应用的关键。在这项工作中，提出了一种新的基于硫化锌半球的改进方法，以显著提高活细胞FTIR成像的空间分辨率。通过将两个高折射率圆顶夹在样品中，在6μm波长下获得了接近2.2μm的横向分辨率，即低于空气中的理论衍射极限，比我们之前使用CaF_2透镜的提高了一倍多(~λ/2.7)。硫化锌圆顶还允许将光谱范围扩展到950厘米−1，而使用CaF2的截止光谱范围为1050厘米−1。与同步辐射光源相结合，MicroFTIR通过约12μm的薄层介质提供了更高的信噪比，从而使脂肪、蛋白质和核酸能够在单个活细胞的细胞核周围得到详细的分布。脂类ν(CH)和ν(C=O)条带清晰显示内质网，ν(PO2−)条带突出核区。这项工作还包括药物(阿霉素)在细胞测量中的演示，以突出这种方法的潜力。图表摘要本文的在线版本(10.1007/s002160181245x)包含补充材料，授权用户可以使用。
FTIR imaging is a label-free, non-destructive method valuably exploited in the study of the biological process in living cells. However, the long wavelength/low spatial resolution and the strong absorbance of water are still key constrains in the application of IR microscopy ex vivo. In this work, a new retrofit approach based on the use of ZnS hemispheres is introduced to significantly improve the spatial resolution on live cell FTIR imaging. By means of two high refractive index domes sandwiching the sample, a lateral resolution close to 2.2 μm at 6 μm wavelength has been achieved, i.e. below the theoretical diffraction limit in air and more than twice the improvement (to ~λ/2.7) from our previous attempt using CaF2 lenses. The ZnS domes also allowed an extended spectral range to 950 cm−1, in contrast to the cut-off at 1050 cm−1 using CaF2. In combination with synchrotron radiation source, microFTIR provides an improved signal-to-noise ratio through the circa 12 μm thin layer of medium, thus allowing detailed distribution of lipids, protein and nucleic acid in the surround of the nucleus of single living cells. Endoplasmic reticula were clearly shown based on the lipid ν(CH) and ν(C=O) bands, while the DNA was imaged based on the ν(PO2−) band highlighting the nucleus region. This work has also included a demonstration of drug (doxorubicin) in cell measurement to highlight the potential of this approach. Graphical abstract The online version of this article (10.1007/s00216-018-1245-x) contains supplementary material, which is available to authorized users.
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影响因子：
1.6
作者：
Carr, GL
通讯作者：
Carr, GL
影响因子：
3.5
作者：
Chan, KLA;Kazarian, SG
通讯作者：
Kazarian, SG
DOI：
10.1016/j.bbamcr.2015.07.018
发表时间：
2015-10-01
影响因子：
5.1
作者：
Fale, Pedro L.;Altharawi, Ali;Chan, K. L. Andrew
通讯作者：
Chan, K. L. Andrew
影响因子：
4.2
作者：
Chan, K. L. Andrew;Kazarian, Sergei G.
通讯作者：
Kazarian, Sergei G.
影响因子：
6.1
作者：
Birarda, G.;Ravasio, A.;Grenci, G.
通讯作者：
Grenci, G.